Phosphoregulation of Arp2/3-dependent actin assembly during receptor-mediated endocytosis.
In both yeast and mammals, endocytic internalization is accompanied by a transient burst of actin polymerization. The yeast protein kinases Prk1p and Ark1p, which are related to the mammalian proteins GAK and AAK1, are key regulators of this process. However, the molecular mechanism(s) by which they regulate actin assembly at endocytic sites have not yet been determined. The Eps15-like yeast protein Pan1p is a Prk1p substrate that is essential for endocytic internalization and for proper actin organization. Pan1p is an Arp2/3 activator and here we show that this activity is dependent on F-actin binding. Mutation of all 15 Prk1p-targeted threonines in Pan1p to alanines mimicked the ark1Delta prk1Delta phenotype, demonstrating that Pan1p is a key Prk1p target in vivo. Moreover, phosphorylation by Prk1p inhibited the ability of Pan1p to bind to F-actin and to activate the Arp2/3 complex, thereby identifying the endocytic phosphoregulation mechanism of Prk1p. We conclude that Prk1p phosphorylation of Pan1p shuts off Arp2/3-mediated actin polymerization on endocytic vesicles, allowing them to fuse with endosomes.


Deep learning guided image-based droplet sorting for on-demand selection and analysis of single cells and 3D cell cultures.
Uncovering the heterogeneity of cellular populations and multicellular constructs is a long-standing goal in fields ranging from antimicrobial resistance to cancer research. Emerging technology platforms such as droplet microfluidics hold the promise to decipher such heterogeneities at ultra-high-throughput. However, there is a lack of methods able to rapidly identify and isolate single cells or 3D cell cultures. Here we demonstrate that deep neural networks can accurately classify single droplet images in real-time based on the presence and number of micro-objects including single mammalian cells and multicellular spheroids. This approach also enables the identification of specific objects within mixtures of objects of different types and sizes. The training sets for the neural networks consisted of a few hundred images manually picked and augmented to up to thousands of images per training class. Training required less than 10 minutes using a single GPU, and yielded accuracies of over 90% for single mammalian cell identification. Crucially, the same model could be used to classify different types of objects such as polystyrene spheres, polyacrylamide beads and MCF-7 cells. We applied the developed method for the selection of 3D cell cultures generated with Hek293FT cells encapsulated in agarose gel beads, highlighting the potential of the technology for the selection of objects with a high diversity of visual appearances. The real-time sorting of single droplets was in-line with droplet generation and occurred at rates up to 40 per second independently of image size up to 480 × 480 pixels. The presented microfluidic device also enabled storage of sorted droplets to allow for downstream analyses.


How flat can a horse be? Exploring 2D approximations of 3D crania in equids.
Quantitative analyses of morphological variation using geometric morphometrics are often performed on 2D photos of 3D structures. It is generally assumed that the error due to the flattening of the third dimension is negligible. However, despite hundreds of 2D studies, few have actually tested this assumption and none has done it on large animals, such as those typically classified as megafauna. We explore this issue in living equids, focusing on ventral cranial variation at both micro- and macro-evolutionary levels. By comparing 2D and 3D data, we found that size is well approximated, whereas shape is more strongly impacted by 2D inaccuracies, as it is especially evident in intra-specific analyses. The 2D approximation improves when shape differences are larger, as in macroevolution, but even at this level precise inter-individual similarity relationships are altered. Despite this, main patterns of sex, species and allometric variation in 2D were the same as in 3D, thus suggesting that 2D may be a source of 'noise' that does not mask the main signal in the data. However, the picture that emerges from this and other recent studies on 2D approximation of 3D structures is complex and any generalization premature. Morphometricians should therefore test the appropriateness of 2D using preliminary investigations in relation to the specific study questions in their own samples. We discuss whether this might be feasible using a reduced landmark configuration and smaller samples, which would save time and money. In an exploratory analysis, we found that in equids results seem robust to sampling, but become less precise and, with fewer landmarks, may slightly overestimate 2D inaccuracies.


[Correction of atherogenic dyslipidemia with honey, pollen and bee bread in patients with different body mass].
To assess efficacy of treatment of patients with atherogenic dyslipidemia (ADL) with beekeeping products (honey, pollen, bee bread). ADL parameters were examined in 157 patients (64 males and 93 females) aged 39 to 72 (mean age 61,7 + 8,5 years) with ADL. Products of beekeeping were given in the absence of allergy and individual resistance to honey, pollen, bee bread. The patients were divided into four groups: patients on hypolipidemic diet only, on diet and honey or pollen, on bee bread, combined treatment - diet, honey, pollen. A significant hypolipidemic effect was registered in patients taking honey in combination with pollen (total cholesterol decreased by 18,3 %, LDLP cholesterol by 23,9 %) and bee bread (total cholesterol decreased by 15,7 %, LDLP cholesterol by 20,5 %). Improvement of blood lipid composition in taking honey and pollen in overweight (body mass index - BMI 25 - 30) and obese (BMI over 30) patients occurs only in loss of body mass.


Outcome of minimally invasive surgical treatment of heartworm caval syndrome in dogs: 42 cases (1999-2007).
To report the outcome of minimally invasive surgical treatment of heartworm caval syndrome in a series of dogs and to provide information on long-term survival of patients with this condition. Retrospective case series. 42 client-owned dogs with a diagnosis of heartworm caval syndrome. Information on history, clinical, laboratory, and diagnostic imaging findings and treatment was obtained from medical records. When possible, additional follow-up information was obtained through telephone interviews with referring veterinarians and owners. Of the 42 dogs with caval syndrome, 21 underwent minimally invasive surgical treatment consisting of transvenous heartworm extraction. Two of the 21 dogs died during the procedure, and after surgery, 4 died. Following induction of anesthesia, heartworms migrated into the distal portion of the pulmonary artery in 1 dog; therefore, extraction was not attempted. Transvenous heartworm extraction was completed successfully in 14 dogs, and all 14 of these dogs were discharged from the hospital. Mean follow-up time in these 14 dogs was 24.4 +/- 17.7 months with a range of 2 to 56 months. At the time of final follow-up, 10 of these 14 dogs had survived at least 18 months and 7 had survived > 24 months. By the end of the study, 1 dog was lost to follow-up and 3 had been euthanatized for unrelated reasons. Results of the study reported here suggest that dogs with caval syndrome that undergo successful transvenous heartworm extraction and survive to discharge have a good long-term prognosis.


Proanthocyanidin-chitosan composite nanoparticles prevent bacterial invasion and colonization of gut epithelial cells by extra-intestinal pathogenic Escherichia coli.
Cranberry proanthocyanidin-chitosan composite nanoparticles (PAC-CHT NPs) were formulated using 2:1, 5:1, 10:1, 15:1 20:1, 25:1, and 30:1 PAC to CHT weight ratio to form round shaped particles. The PAC-CHT NPs were characterized by size, polydispersity, surface charge, morphology, and PAC content. PAC-CHT NPs bioactivity was measured by agglutination of extra-intestinal pathogenic Escherichia coli (ExPEC) and inhibition of gut epithelial cell invasion by ExPEC. Results indicate that by increasing the PAC to CHT ratio 10:1 to 30:1 formed stable nanoparticles with diameters of 122.8 to 618.7 nm, a polydispersity index of approximated 0.4 to 0.5, and a zeta potential of 34.5 to 54.4 mV. PAC-CHT NPs ratio 30:1 agglutinated ExPEC and decreased the ability of ExPEC to invade epithelial cells in a dose-dependent manner. PAC-CHT NPs ratio 10:1 to 30:1 form stable, round-shaped, and bioactive nanoparticles for potential applications in the treatment of ExPEC bacterial infections.


Cyclooxygenase selectivity of nonsteroidal anti-inflammatory drugs in canine blood.
To evaluate cyclooxygenase (COX) selectivity of several nonsteroidal anti-inflammatory drugs (NSAID) in canine blood in vitro. 11 healthy adult male hound crosses. 9 NSAID were studied at 5 concentrations. Thromboxane B2 (TxB2) was assayed as a measure of COX-1 activity in clotted blood. Prostaglandin E2 (PGE2) was assayed as a measure of COX-2 activity in heparinized, lipopolysaccharide (LPS)-stimulated blood. All assays were competitive ELISA tests. Cyclooxygenase selectivity was expressed as a ratio of the concentration of an NSAID that inhibited 50% of the activity (IC50) of COX-1 to the IC50 of COX-2. A separate ratio of the concentration that inhibited 80% of COX activity (IC80) was also determined. A ratio of < 1.0 indicated selectivity for COX-1, whereas a ratio of > 1.0 indicated COX-2 selectivity. Ketoprofen, aspirin, and etodolac were COX-1 selective. Piroxicam, meloxicam, and carprofen had COX-2 selectivity. The IC50 and IC80 values were similar for most NSAID. This methodology provides repeatable data from individual dogs and is comparable to results of previous in vitro and ex vivo models. Findings are also consistent with those of canine studies performed in vivo, suggesting that this is a viable in vitro assessment of the COX selectivity of NSAID in dogs.


Methylguanine DNA methyltransferase-mediated drug resistance-based selective enrichment and engraftment of transplanted stem cells in skeletal muscle.
Cell replacement therapy using stem cell transplantation holds much promise in the field of regenerative medicine. In the area of hematopoietic stem cell transplantation, O(6)-methylguanine-DNA methyltransferase MGMT (P140K) gene-mediated drug resistance-based in vivo enrichment strategy of donor stem cells has been shown to achieve up to 75%-100% donor cell engraftment in the host's hematopoietic stem cell compartment following repeated rounds of selection. This strategy, however, has not been applied in any other organ system. We tested the feasibility of using this MGMT (P140K)-mediated enrichment strategy for cell transplantation in skeletal muscles of mice. We demonstrate that muscle cells expressing an MGMT (P140K) drug resistance gene can be protected and selectively enriched in response to alkylating chemotherapy both in vitro and in vivo. Upon transplantation of MGMT (P140K)-expressing male CD34(+ve) donor stem cells isolated from regenerating skeletal muscle into injured female muscle treated with alkylating chemotherapy, donor cells showed enhanced engraftment in the recipient muscle 7 days following transplantation as examined by quantitative-polymerase chain reaction using Y-chromosome specific primers. Fluorescent in situ hybridization analysis using a Y-chromosome paint probe revealed donor-derived de novo muscle fiber formation in the recipient muscle 14 days following transplantation, with approximately 12.5% of total nuclei within the regenerated recipient muscle being of donor origin. Following engraftment, the chemo-protected donor CD34(+ve) cells induced substantial endogenous regeneration of the chemo-ablated host muscle that is otherwise unable to self-regenerate. We conclude that the MGMT (P140K)-mediated enrichment strategy can be successfully implemented in muscle.


Sun exposure related methylation in malignant and non-malignant skin lesions.
We investigated the aberrant promoter methylation status of 12 genes in skin lesions, both malignant (basal cell carcinomas (BCCs), n=68 and squamous cell carcinomas (SCCs), n=35) and non-malignant (tags, n=58) skin lesions and compared the results of lesions from sun exposed (SE) and sun protected (SP) regions. Methylation was studied using a methylation specific PCR (MSP) and methylation of CDH1 was also measured using a semi-quantitative fluorescence based real-time MSP method. The methylation index (MI) was calculated as the methylated fraction of the genes examined. In this report, we found high frequencies of methylation of several known or suspected tumor suppressor genes in tags and skin cancers. Among the 12 genes, for the cadherin genes CDH1 and CDH3 and for two of the laminin 5 encoding genes LAMA3 and LAMC2 methylation frequencies greater than 30% were noted in one or more specimen types. We investigated whether methylation was tumor related. Surprisingly, the differences in the methylation profile of genes among the three specimen types were modest, and the MI, indicators of overall methylation frequencies, was nearly identical. However, significant differences were noted in the frequencies of methylation among the three specimen types for the genes RASSF1A (P=0.002), CDH1 (P=0.007) and one or more of three CAD genes (P=0.02). Methylation was highly significantly related to sun exposure, and sun protected specimens had little or no methylation. As methylation of CDH1 was completely SE specific we analyzed all the skin samples using a semi-quantitative real-time PCR assay for the CDH1 gene. The concordance between standard MSP and real-time MSP for all the samples (n=161) was 75% (P<0.0001). While weak signals were detected in the SP samples by real time PCR, the differences between SE and SP specimens were 148 fold for tags and 390 fold for BCCs. These differences were highly significant (P<0.0001). These findings suggest that methylation commences in UV exposed skin at a relatively early age and occurs in skin prior to the onset of recognizable preneoplastic changes.


Anti-Glomerular Basement Membrane Disease Combined with IgA Nephropathy Complicated with Reversible Posterior Leukoencephalopathy Syndrome: An Unusual Case.
Anti-glomerular basement membrane disease (anti-GBM disease) is an autoimmune glomerulonephritis disease that is characterized by IgG linear deposition along the non-collagen domain of a3 chains of type IV collagen on the GBM. Although anti-GBM disease accompanied with IgA linear deposition along GBMs was discussed previously in some papers, anti-GBM disease combined with IgA granular deposition in the mesangial area, especially complicated with reversible posterior leukoencephalopathy syndrome (RPLS), was rarely reported. RPLS is usually caused by hypertensive encephalopathy, renal decompensation, fluid retention, and adverse effects of immunosuppressive drugs. A male patient with the chief complaints of headache, gross hematuria, and nocturia was referred to our hospital. Based on renal biopsy, the diagnosis was finally confirmed as anti-GBM disease combined with IgA nephropathy and, the patient received comprehensive treatment, including cyclophosphamide (CTX), which led to symptom improvement. Two days after the third impulse CTX was given, he suddenly experienced headache and dizziness, which eventually developed into a tonic-clonic seizure. RPLS was identified by cranial magnetic resonance imaging (MRI) with reversible neuroimaging. After diazepam and antihypertension management, seizures were controlled. RPLS, a neurological complication, was found in anti-GBM disease with IgA nephropathy during our immunosuppressants therapy for the first time. It is worth paying more attention to patients with rapidly progressive glomerulonephritis (RPGN), as they might be complicated with RPLS during intravenous administration of CTX and methylprednisolone. We suggest the neuroimaging be examined as soon as the seizure happens.


A multicenter case-control study of diet and lung cancer among non-smokers.
We have examined the role of dietary patterns and specific dietary nutrients in the etiology of lung cancer among non-smokers using a multicenter case-control study. 506 non-smoking incident lung cancer cases were identified in the eight centers along with 1045 non-smoking controls. Dietary habits were assessed using a quantitative food-frequency questionnaire administered by personal interview. Based on this information, measures of total carotenoids, beta-carotene and retinol nutrient intake were estimated. Protective effects against lung cancer were observed for high consumption of tomatoes, (odds ratio (OR) = 0.5; 95% confidence interval (CI) 0.4-0.6), lettuce (OR = 0.6; 95% CI 0.3-1.2), carrots (OR = 0.8; 95% CI 0.5-1.1), margarine (OR = 0.7; 95% CI 0.5-0.8) and cheese (OR = 0.7; 95% CI 0.5-1.0). Only weak protective effects were observed for high consumption of all carotenoids (OR = 0.8; 95% CI 0.6-1.0), beta-carotene (OR = 0.8; 95% CI 0.6-1.1) and retinol (OR = 0.9; 95% CI 0.7-1.1). Protective effects for high levels of fruit consumption were restricted to squamous cell carcinoma (OR = 0.7; 95% CI 0.4-1.2) and small cell carcinoma (OR = 0.7; 95% CI 0.4-1.2), and were not apparent for adenocarcinoma (OR = 0.9; 95% CI 0.6-1.3). Similarly, any excess risk associated with meat, butter and egg consumption was restricted to squamous and small cell carcinomas, but was not detected for adenocarcinomas. This evidence suggests that the public health significance of increasing vegetable consumption among the bottom third of the population would include a reduction in the incidence of lung cancer among lifetime non-smokers by at least 25%, and possibly more. A similar protective effect for increased fruit consumption may be present for squamous cell and small cell lung carcinomas.


A 5-year clinical study of indirect and direct resin composite and ceramic inlays.
The aim of this investigation was to evaluate the clinical performance of 4 types of tooth-colored inlays. Fifteen direct ceramic inlays (Cerec Cos 2.0), 15 direct resin composite inlays (Brilliant Direct Inlay), 14 indirect ceramic inlays (Vita Dur N), and 14 indirect resin composite inlays (Estilux) were made in 37 patients, according to the manufacturers' instructions. The inlays were evaluated 1 week (baseline) and 6, 12, 36, 48, and 60 months after cementation (modified CDA Quality Evaluation System). Two Vita Dur N inlays fractured after 1 and 4 years in function, and one Cerec inlay fractured after 4.5 years. Two Brilliant DI inlays needed replacement because of secondary caries (after 1 and 5 years), and one inlay (Estilux) needed replacement due to persisting hypersensitivity. Three inlays (1 Estilux and 2 Brilliant DI) were repaired due to chipping or minor fractures. During the observation period, the surface texture of Brilliant DI and Vita Dur N inlays became significantly rougher. After 5 years, the Estilux inlays had significantly lower ratings for morphology compared to baseline ratings. In general, the occlusal marginal adaptation did not show further disintegration of the luting cement after 1 year. Eighty-eight percent of the inlays were in function after 5 years. No significant differences were revealed among the survival rates of the different types of inlays.


Evaluation of Fungicides for the Management of Botryosphaeria Canker of Grapevines.
The family Botryosphaeriaceae comprises a number of species that are associated with the dieback disease of grapevine (Vitis vinifera), referred to as Botryosphaeria canker. To date, there are few effective agents available for the management of this disease. In this study, fungicides were evaluated for controlling the disease using a combination of in vitro tests and field trials. Twenty fungicides registered for use on other diseases in Australian viticulture were tested in vitro for their effect on mycelial growth of four species within the Botryosphaeriaceae. The concentrations of fungicide at which 50% of mycelial growth is inhibited (EC50 values) were significantly affected both by fungicide and isolate (P < 0.001). Differences in sensitivities of the four species to the fungicides were negligible (0.41 to 0.59 mg/liter). The most effective fungicides were fludioxonil, carbendazim, fluazinam, tebuconazole, flusilazole, penconazole, procymidone, iprodione, myclobutanil, and pyraclostrobin, for which EC50 values were <1.0 mg/liter. These fungicides were evaluated under field conditions, in addition to the pruning wound protectants Bacseal Super, Garrison, and ATCS tree wound dressing, as well as the biological control agent Vinevax. In field trials, carbendazim (Bavistin), fluazinam (Shirlan), tebuconazole (Folicur), Garrison, and ATCS tree wound dressing applied to freshly cut pruning wounds were the most effective and reduced infection by Diplodia seriata and D. mutila by 41 to 65%. These results suggest that the occurrence of Botryosphaeria canker on grapevines may be reduced via treatment of pruning wounds with selected fungicides as soon as possible after pruning.


Indoor and peridomestic transmission of American cutaneous leishmaniasis in northwestern Argentina: a retrospective case-control study.
A case-control study was carried out during 1990-1994 to identify risk factors associated with American cutaneous leishmaniasis (ACL) in Santiago del Estero, Argentina. The study subjects consisted of 171 cases and 308 controls matched by age, sex, and place of residence. The analysis was performed by conditional logistic regression. Risk factors found to be significantly associated with ACL were related to indoor transmission (few rooms in the house, dirt floor, and a permanent opening in lieu of a window); peridomestic transmission (presence of a pond or woodland within 150 m of the house and an agricultural area within 200 m of the house); and human behavior (sleeping in the backyard, collecting water, bathing, and performing agricultural activities). Most transmission appears to have occurred indoors and in the peridomicile. These environments should be included in further research and control policies.


Animal-borne acoustic transceivers reveal patterns of at-sea associations in an upper-trophic level predator.
Satellite telemetry data have substantially increased our understanding of habitat use and foraging behaviour of upper-trophic marine predators, but fall short of providing an understanding of their social behaviour. We sought to determine whether novel acoustic and archival GPS data could be used to examine at-sea associations among grey seals (Halichoerus grypus) during the fall foraging period. Fifteen grey seals from Sable Island, Canada were deployed with Vemco Mobile Transceivers and Satellite-GPS transmitters in October 2009, 13 of which were recaptured and units retrieved 79 ± 2.3 days later during the following breeding season, December 2009-January 2010. An association between two individuals was defined as a cluster of acoustic detections where the time between detections was <30 min. Bathymetry, travel rate, and behavioural state (slow and fast movement) were determined for each GPS archival point (3.7 ± 0.1 locations recorded per hour). Behavioural state was estimated using a hidden Markov model. All seals had been involved in associations with other instrumented seals while at sea, with a total of 1,872 acoustic detections recorded in 201 associations. The median number of detections per association was 3 (range: 1-151) and the median duration of an association was 0.17 h (range: <0.1-11.3 h). Linear mixed-effects models showed that associations occurred when seals were exhibiting slow movement (0.24 ± 0.01 ms(-1)) on shallow (53.4 ± 3.7 m) offshore banks where dominant prey is known to occur. These results suggest the occurrence of short-term associations among multiple individuals at foraging grounds and provide new insights into the foraging ecology of this upper-trophic marine predator.


Hyperbaric Oxygen Effects on Depression-Like Behavior and Neuroinflammation in Traumatic Brain Injury Rats.
The aim of this study was to determine whether hyperbaric oxygen (HBO) therapy causes attenuation of traumatic brain injury (TBI)-induced depression-like behavior and its associated anti-neuroinflammatory effects after fluid percussion injury. Anesthetized male Sprague-Dawley rats were divided into 3 groups: sham operation plus normobaric air (NBA) (21% oxygen at 1 absolute atmosphere [ATA]), TBI plus NBA, and TBI plus HBO (100% oxygen at 2.0 ATA). HBO was applied immediately for 60 min/d after TBI for 3 days. Depression-like behavior was tested by a forced swimming test, motor function was tested by an inclined plane test, and infarction volume was tested by triphenyltetrazolium chloride (TTC) staining on days 4, 8, and 15. Neuronal apoptosis (terminal deoxynucleotidyl transferase dUTP nick-end labeling assay), microglial (marker OX42) activation, and tumor necrosis factor (TNF)-α expression in microglia in the hippocampus CA3 were measured by immunofluorescence methods. Compared with the TBI controls, without significant changes in TTC staining or in the motor function test, TBI-induced depression-like behavior was significantly attenuated by HBO therapy by day 15 after TBI. Simultaneously, TBI-induced neuronal apoptosis, microglial (marker OX42) activation, and TNF-α expression in the microglia in the hippocampus CA3 were significantly reduced by HBO. Our results suggest that HBO treatment may ameliorate TBI-induced depression-like behavior in rats by attenuating neuroinflammation, representing one possible mechanism by which depression-like behavior recovery might occur. We also recommend HBO as a potential treatment for TBI-induced depression-like behavior if early intervention is possible.


Personality traits as predictors of early alcohol inebriation among young adolescents: Mediating effects by mental health and gender-specific patterns.
The aim of this study was to predict alcohol inebriation and mental health (internalizing and externalizing problems plus well-being), and potential gender-specific patterns among young adolescents, by a biopsychosocial model of personality traits. Self-reported data from 853 adolescents (479 girls) in Sweden, aged 13-15 years, from the Longitudinal Research on Development In Adolescence (LoRDIA) program were used. Predictions from personality to inebriation and mediating effects of mental health were estimated by means of logistic regression and generalized structural equation modelling. Separated gender analyses were performed throughout the study to reveal potential gender-specific patterns. Externalizing problems, Novelty Seeking and Cooperativeness had independent effects on alcohol inebriation for both genders as well as Harm Avoidance among girls and Internalizing problems among boys. Novelty Seeking and Self-Directedness had indirect effects through externalizing problems and Harm Avoidance and Self-Directedness had indirect effects through internalizing problems for boys. Self-directedness showed an indirect effect through externalizing problems for girls. The combination of an immature character (low Self-directedness and Cooperativeness) with an extreme temperament profile (high Novelty Seeking and low Harm Avoidance) was a predictor of inebriation across gender, both directly and indirectly through mental health. This study contributes with valuable information about gender-specific considerations when developing and conducting preventative interventions targeting psychological risk and resilience factors for early alcohol inebriation among young adolescents.


Percutaneous Driveline Fracture After Implantation of the HeartMate II Left Ventricular Assist Device: How Durable is Driveline Repair?
Durability of the percutaneous driveline is critical for the optimal long-term support of patients after left ventricular assist device (LVAD) implantation. There are no data specifically examining the durability of major repair for driveline fracture or its effect on patient outcomes. Between May 2004 and August 2014, 560 patients underwent implantation with the HeartMate II LVAD at our respective institutions. Median age at LVAD implantation was 59 years (range 18-82 years) and 465 patients (83%) were male. The indication for LVAD implantation was bridge to transplant (BTT) in 296 patients (53%). Follow-up was available in all early survivors (n = 519) for a total of 940 patient-years of support (median 1.1 years, max. 10.4 years). Percutaneous driveline fracture was identified in 17 patients (3.2%), eight of whom were implanted as destination therapy. Median time from LVAD implantation to driveline fracture was 1.3 years (range, from 4 months to 3.8 years). Two of these patients underwent device exchange as primary treatment, while 15/17 (88%) underwent repair. Three of these 15 patients required a driveline reintervention, including device exchange (n = 1), rerepair (n = 1), and ungrounded cables (n = 1). Median time of support after driveline repair was 10 months (range from 3 months to 5.4 years). There were no late deaths after driveline repair during the follow-up period with 14/15 patients (93%) active on support and one having undergone transplant. The incidence of percutaneous driveline fracture after HeartMate II LVAD implantation is low (3.9%). The majority of driveline repairs for driveline fracture are durable with reintervention required in a minority of patients. There was no adverse effect of driveline fracture requiring repair on late outcome noted in this series with all patients either active on support or transplanted at last follow-up.


Emergency treatment of acute asthma with albuterol metered-dose inhaler plus holding chamber: how often should treatments be administered?
To determine the optimal treatment interval for administering albuterol metered-dose inhaler (MDI) with a holding chamber to patients presenting to the emergency department (ED) with acute asthma. Prospective, randomized, double-blind study. EDs of two affiliated teaching hospitals in the Bronx, NY. One hundred adult patients with acute asthma and FEV1 <60% predicted of normal. At entry (T=0 min), eligible patients all openly received inhaled albuterol (six puffs) via MDI with a spacer. Subsequently, in a double-blind fashion, they received six puffs of albuterol or placebo with new MDIs and spacers at 30, 60, and 90 min such that group 1 (n=34) received albuterol every 30 min, group 2 (n=33) every 60 min, and group 3 (n=33) at 120 min only. FEV1 and vital signs were measured at T=0 and at 15, 30, 60, 90, and 120 min following initial treatment. Potassium levels were measured at T=0 and 120 min. Adverse events, the use of additional inhaled beta-agonists or systemic corticosteroids, and hospitalization rates were recorded. At T=0, the groups did not differ in age, FEV1, or prescribed asthma medications. All groups showed significant improvement in FEV1 (p<0.05; T=120 vs 0 min). The conditions of groups 1 and 2 improved significantly more than those of group 3, but did not differ compared to each other. The mean+/-SEM change in FEV1 (T=120 vs 0 min) was 0.993+/-0.108, 0.858+/-0.135, and 0.321+/-0.056 L, respectively, for the three groups. Separate analysis for patients with FEV1% <40% or >40% predicted showed similar results. However, patients who initially were low responders to albuterol treatment (<15 percentage point increase at 15 min) improved significantly with 30-min treatments compared to the other two treatment regimens. Patients who initially responded with >15 percentage point increase in FEV1 at 15 min following initial albuterol inhalation benefited equally from 30- or 60-min treatments compared to 120 min. Potassium levels did not change significantly during the study. Adverse events and hospitalization rates were equivalent. After the conclusion of the study, group 3 patients required a greater number of beta-agonist treatments prior to eventual discharge from the ED. For acute asthma, albuterol MDI with a holding chamber can be given optimally at 60-min intervals with minimal adverse effects for the majority of patients. However, patients who initially demonstrate a low or poor bronchodilator response to albuterol should be given subsequent treatments at 30-min intervals. This will optimize care and conserve resources for patients who will benefit the most.


Electrically induced and voluntary activation of physiologic muscle pump: a comparison between spinal cord-injured and able-bodied individuals.
To evaluate the central haemodynamic responses during position changes from supine to sitting and during 30 min of standing between able-bodied and spinal cord-injured subjects. Also to assess the effects of the physiologic muscle pump in both groups during 30 min of standing. A repeated measure design. Both groups were tested on two different days under two conditions of 30 min of stationary standing and 30 min of dynamic standing (voluntary activation of the lower leg muscles in able-bodied and FES-induced activation of these muscles in spinal cord injured). The order of testing was random. Rehabilitation hospital. Fifteen healthy able-bodied and 14 healthy spinal cord-injured subjects. Stroke volume, cardiac output, heart rate, systolic blood pressure, diastolic blood pressure, mean arterial pressure and total peripheral resistance during supine-pre sitting, sitting-pre standing and during 30 min of standing. Significant reductions (p < 0.05) in systolic blood pressure, diastolic blood pressure and mean arterial pressure were found when spinal cord-injured subjects moved from sitting to standing during stationary standing; these values were maintained during dynamic standing. These values were maintained during both standing sessions in able-bodied subjects. During 30 min of stationary standing, there were significant reductions in stroke volume, cardiac output in both able-bodied and spinal cord-injured while their total peripheral resistance increased (p < 0.05). During 30 min of dynamic standing, both groups maintained their haemodynamics at pre-standing values with the exception of significant reduction in stroke volume at 30 min of standing. FES-induced activation of the physiologic muscle pump during change in position from sitting to standing prevented orthostatic hypotension in spinal cord-injured subjects. During standing it had equal or even greater effect on improving blood circulation when compared with voluntary activation in able-bodied subjects. The use of FES during standing and tilting in spinal cord-injured individuals may prevent orthostatic hypotension and circulatory hypokinesis and improve tolerance to tilting and standing.


On the time course of visual word recognition: an event-related potential investigation using masked repetition priming.
The present study used event-related potentials (ERPs) to examine the time course of visual word recognition using a masked repetition priming paradigm. Participants monitored target words for occasional animal names, and ERPs were recorded to nonanimal critical items that were full repetitions, partial repetitions, or unrelated to the immediately preceding masked prime word. The results showed a strong modulation of the N400 and three earlier ERP components (P150, N250, and the P325) that we propose reflect sequential overlapping steps in the processing of printed words.


FOREST ECOLOGY. Pervasive drought legacies in forest ecosystems and their implications for carbon cycle models.
The impacts of climate extremes on terrestrial ecosystems are poorly understood but important for predicting carbon cycle feedbacks to climate change. Coupled climate-carbon cycle models typically assume that vegetation recovery from extreme drought is immediate and complete, which conflicts with the understanding of basic plant physiology. We examined the recovery of stem growth in trees after severe drought at 1338 forest sites across the globe, comprising 49,339 site-years, and compared the results with simulated recovery in climate-vegetation models. We found pervasive and substantial "legacy effects" of reduced growth and incomplete recovery for 1 to 4 years after severe drought. Legacy effects were most prevalent in dry ecosystems, among Pinaceae, and among species with low hydraulic safety margins. In contrast, limited or no legacy effects after drought were simulated by current climate-vegetation models. Our results highlight hysteresis in ecosystem-level carbon cycling and delayed recovery from climate extremes.


Age-specific survival and reproductive probabilities: evidence for senescence in male fallow deer (Dama dama).
Survival and reproduction are key features in the evolution of life-history strategies. In this study, we use capture-mark-resighting and multi-state models to examine survival senescence and reproductive senescence in six successive cohorts of fallow bucks that were studied for 16 years. We found that the overall age-specific survival probabilities of males were highly variable and the best-fitting model revealed that fallow bucks have four life-history stages: yearling, pre-reproductive, prime-age and senescent. Pre-reproductive males (2 and 3 years old) had the highest survival. Survival declined sharply after the age of 9 years, indicating that senescence had begun. When we considered reproducing and non-reproducing males separately, there was no evidence of senescence in the former, and steadily decreasing survival after the onset of social maturity in the latter. Reproduction probability also declined in older males, and thus we provide very strong evidence of senescence. Reproducers had a greater chance of reproducing again in the following year than non-reproducers. Furthermore, there were differences in the survival probabilities, with reproducers consistently surviving better than non-reproducers. In our study population, reproducers allocate more to the effort to reproduce than non-reproducers. Therefore our results indicate the generally higher phenotypic quality of reproducing males. These results, along with earlier studies on the same population, could indicate positive relationships between fitness correlates.


Protein S exacerbates alcoholic hepatitis by stimulating liver natural killer T cells.
Alcohol consumption is a major cause of liver injury but the mechanisms are not completely understood. Protein S (PS) is an anticoagulant glycoprotein with multiple functions. The role of PS in liver injury is unknown. This study investigated the role of PS in acute alcoholic hepatitis. A mouse overexpressing human PS (hPS-TG) was generated in which acute hepatitis was induced by intraperitoneal injection of ethanol. The levels of serum liver enzymes and liver tissue inflammatory cytokines and the degree of hepatic steatosis were significantly increased in hPS-TG mice treated with ethanol compared with ethanol-treated wild type (WT) mice. Cell expansion, activation and inhibition of apoptosis were significantly augmented in natural killer T (NKT) cells from hPS-TG mice compared with WT mice. Liver mononuclear cells from hPS-TG mice express higher levels of inflammatory cytokines than those from WT mice after stimulation with a specific stimulant of NKT cells in vitro. In a co-culture system of hepatocytes and NKT cells, the effects of PS on ethanol-mediated cell injury were suppressed by a CD1d neutralizing antibody. Alcoholic liver injury was significantly improved in mice pre-treated with PS siRNA and anti-protein S antibody compared with control mice. Patients with alcoholic hepatitis showed significantly increased plasma PS levels and enhanced liver expression of PS and CD1d compared with controls. The results of this study suggest that PS exacerbates acute alcoholic hepatitis by inhibiting apoptosis of activated NKT cells.


Cryptosporidium infection causes undernutrition and, conversely, weanling undernutrition intensifies infection.
Cryptosporidium parvum is a leading pathogen in children in developing countries. To investigate whether early postnatal malnutrition leads to heavier C. parvum infections, we assessed intestinal adaptation and parasite load in suckling mice during the first 2 wk of life, analogous to the first postnatal yr in humans. Undernutrition was induced by daily C57BL6J pup separation from lactating dams. Half of the pups were separated daily, for 4 hr on day 4, 8 hr on day 5, and for 12 hr from day 6 until day 14. On day 6, each pup received an oral inoculum of 10(5) to 10(7) parasites in 10-25 microl of PBS. Littermate controls received PBS alone. Stools were assessed from days 8, 11, and 14 for oocyst counts. Mice were killed on day 14, 8 days postinoculation, at the peak of the infection. Ileal and colon segments were obtained for histology, real-time and reverse transcriptase PCR, and immunoassays. Villus and crypt lengths and cross-sectional areas were also measured. Undernourished and nourished mice infected with excysted 10(6) or 10(7) oocysts exhibited the poorest growth outcomes compared with their uninfected controls. Nourished 10(6)-infected mice had comparable weight decrements to uninfected undernourished mice. Body weight and villi were additively affected by malnutrition and cryptosporidiosis. Hyperplastic crypts and heavier inflammatory responses were found in the ilea of infected malnourished mice. Undernourished infected mice exhibited greater oocyst shedding, TNF-alpha and IFN-gamma intestinal levels, and mRNA expression compared to nourished mice infected with either 10(5) or 10(6) oocysts. Taken together, these findings show that Cryptosporidium infection can cause undernutrition and, conversely, that weanling undernutrition intensifies infection and mucosal damage.


Prostaglandin E2 effects on corneal endothelial cyclic adenosine monophosphate synthesis and cell shape are mediated by a receptor of the EP2 subtype.
Corneal endothelial cells synthesize prostaglandin E2 (PGE2), and this synthesis is necessary for the maintenance of the normal polygonal shape of these cells. A series of experiments was done to examine the receptor-effector mechanism responsible for PGE2-mediated effects on cultured rabbit corneal endothelium. When challenged with exogenous PGE2, endothelial cells synthesized cyclic adenosine monophosphate (AMP) in a dose-dependent manner, and this synthesis was not antagonized by AH6809. The synthetic agonist 11-deoxy-PGE1, but not sulprostone, stimulated increased cyclic AMP synthesis. The pharmacologic profile of the endothelial PGE2 receptor is therefore consistent with that of an EP2 receptor linked to activation of adenylate cyclase. The prostaglandin agonists were also tested for their ability to prevent cellular elongation in response to indomethacin. The PGE2, 11-deoxy-PGE1, and 16,16-dimethyl PGE1 prevented elongation, but sulprostone and PGF2 alpha did not. The authors conclude that rabbit corneal endothelium in culture expresses a specific PG receptor of the EP2 subtype which is coupled to cyclic AMP synthesis and is involved in the regulation of cell shape.


Ecotypic variation in the context of global climate change: revisiting the rules.
Patterns of ecotypic variation constitute some of the few 'rules' known to modern biology. Here, we examine several well-known ecogeographical rules, especially those pertaining to body size in contemporary, historical and fossil taxa. We review the evidence showing that rules of geographical variation in response to variation in the local environment can also apply to morphological changes through time in response to climate change. These rules hold at various time scales, ranging from contemporary to geological time scales. Patterns of body size variation in response to climate change at the individual species level may also be detected at the community level. The patterns underlying ecotypic variation are complex and highly context-dependent, reducing the 'predictive-power' of ecogeographical rules. This is especially true when considering the increasing impact of human activities on the environment. Nonetheless, ecogeographical rules may help interpret the likely influences of anthropogenic climate change on ecosystems. Global climate change has already influenced the body size of several contemporary species, and will likely have an even greater impact on animal communities in the future. For this reason, we highlight and emphasise the importance of museum specimens and the continued need for documenting the earth's biological diversity.


Prospective trials of minor surgical procedures and high-fibre diet for haemorrhoids.
Two hundred and sixteen consecutive patients seen for the first time with symptomatic haemorrhoids entered trials of conservative and minor surgical treatment. They were divided into two groups according to their pretreatment maximal anal pressure. Patients with pressures of 100 cm H2O (73.7 mm Hg) or more (108 patients) were treated by anal dilatation (37), sphincterotomy (34), or high-fibre diet (37). Four and 12 months later anal dilatation had produced significantly better results than sphincterotomy or diet. Furthermore, anal dilatation was the only treatment associated with a significant reduction in anal pressure at four and 12 months. Patients with pressures under 100 cm H2O (108 patients) were treated by rubber-band ligation (35), cryosurgery (36), or diet (37). Four and 12 months later significantly more patients were improved by rubber-band ligation than by cryosurgery or diet. These results suggest that haemorrhoids in patients with excessive activity of the internal anal sphincter are best treated by anal dilatation and that in all other patients rubber-band ligation is the treatment of choice.


Critical period for styrene ototoxicity in the rat.
The current experiments were undertaken to determine whether or not styrene-induced hearing loss in the rat depends more on the existence of a critical period between 14 and 21 weeks of age than on body weight. For these purposes, two experiments were carried out with mature Long-Evans rats. In the first experiment, two groups of 5-month old rats, but having different body weight (slim: 314 g vs. fat: 415 g) were exposed to 700 ppm styrene for 4 consecutive weeks, 5 days per week, 6 hours per day. In the second experiment, two groups of rats having the same weight: 345 g, but different ages (14- vs. 21- week old) were exposed to styrene in strictly identical experimental conditions. Auditory sensitivity was tested by recording evoked potentials from the inferior colliculus. Surface preparations of the organ of Corti were also performed to complete the investigation. At the end of the six week recovery period following the styrene exposure, a 7 dB permanent threshold shift (PTS) was obtained with the same age animals regardless of the body weight. Consequently, weight was not a major factor in styrene-induced hearing loss. Age was a more critical factor in determining higher sensitivity to styrene. Indeed, the three months old group had 23.5 dB PTS, whereas the five months old group had only a 7.7 dB PTS at 16 kHz. Thus, a 15 dB difference of PTS was obtained between the rats having the same weight but different age. While the weight does not play a major role in styrene ototoxicity, there is a critical period whose duration lasts more than three months and for which the susceptibility to styrene is enhanced.


Alginate-loaded liposomes can protect encapsulated alkaline phosphatase functionality when exposed to gastric pH.
The present study has evaluated the potential of alginate-loaded liposomes as a vehicle for the oral delivery of bioactive proteins. The vesicles were prepared from the phospholipid dipalmatoyl phosphatidylcholine using a simple dry film hydration technique. Alkaline phosphatase (ALP) was used as a model bioactive protein, which was encapsulated in alginate-loaded liposomes and conventional liposomes. These vesicles were subject to physical characterization and stability analysis at varying pH values. The enzymatic activity of the ALP was evaluated following exposure to simulated gastric pH. The alginate-loaded liposomes were typically of the order of 10 microm; however, there was evidence of vesicle aggregation thought to be due to alginate present on the surface of the vesicles. The typical size of the aggregated vesicles was approximately 30 microm. The enzyme activity of APL following 2 h of exposure to simulated gastric pH was maintained at a significantly higher level when encapsulated in the alginate-loaded liposomes as compared to ALP loaded in conventional liposomes [80% as compared to 55% (p < 0.05), respectively].


Quantitative analysis of adsorbed proteins by X-ray photoelectron spectroscopy.
Protein adsorption at solid surfaces is central to many phenomena of medical and technological interest. The determination of the amount of protein attached to the surface is a critical measurement performed by using a wide range of methods. X-ray photoelectron spectroscopy (XPS) is able to provide a straightforward quantitative analysis of the amount of protein adsorbed as an overlayer on a material surface. While XPS is commonly employed to assess qualitatively the amount of adsorbed protein, this is usually expressed in terms of the elemental fraction (or at. %) of nitrogen calculated using an assumption of depth homogeneity despite the fact that this does not linearly scale with the amount of protein. In this paper, we have shown that thicknesses derived from XPS data linearly correlated with spectroscopic ellipsometry data on the same samples with a scatter of 10%. A straightforward equation to convert the concentration of nitrogen from XPS into an equivalent thickness of a protein film is presented. We highlight some discrepancies in the absolute thicknesses determined by XPS and ellipsometry on dried films and quartz crystal microbalance on wet films, which appear likely to result from the inclusion of a contribution from water in the latter two techniques.


Early fibrinogen concentrate therapy for major haemorrhage in trauma (E-FIT 1): results from a UK multi-centre, randomised, double blind, placebo-controlled pilot trial.
There is increasing interest in the timely administration of concentrated sources of fibrinogen to patients with major traumatic bleeding. Following evaluation of early cryoprecipitate in the CRYOSTAT 1 trial, we explored the use of fibrinogen concentrate, which may have advantages of more rapid administration in acute haemorrhage. The aims of this pragmatic study were to assess the feasibility of fibrinogen concentrate administration within 45 minutes of hospital admission and to quantify efficacy in maintaining fibrinogen levels ≥ 2 g/L during active haemorrhage. We conducted a blinded, randomised, placebo-controlled trial at five UK major trauma centres with adult trauma patients with active bleeding who required activation of the major haemorrhage protocol. Participants were randomised to standard major haemorrhage therapy plus 6 g of fibrinogen concentrate or placebo. Twenty-seven of 39 participants (69%; 95% CI, 52-83%) across both arms received the study intervention within 45 minutes of admission. There was some evidence of a difference in the proportion of participants with fibrinogen levels ≥ 2 g/L between arms (p = 0.10). Fibrinogen levels in the fibrinogen concentrate (FgC) arm rose by a mean of 0.9 g/L (SD, 0.5) compared with a reduction of 0.2 g/L (SD, 0.5) in the placebo arm and were significantly higher in the FgC arm (p < 0.0001) at 2 hours. Fibrinogen levels were not different at day 7. Transfusion use and thromboembolic events were similar between arms. All-cause mortality at 28 days was 35.5% (95% CI, 23.8-50.8%) overall, with no difference between arms. In this trial, early delivery of fibrinogen concentrate within 45 minutes of admission was not feasible. Although evidence points to a key role for fibrinogen in the treatment of major bleeding, researchers need to recognise the challenges of timely delivery in the emergency setting. Future studies must explore barriers to rapid fibrinogen therapy, focusing on methods to reduce time to randomisation, using 'off-the-shelf' fibrinogen therapies (such as extended shelf-life cryoprecipitate held in the emergency department or fibrinogen concentrates with very rapid reconstitution times) and limiting the need for coagulation test-based transfusion triggers. ISRCTN67540073 . Registered on 5 August 2015.


Cloning and analysis of two new isoforms of multifunctional Ca2+/calmodulin-dependent protein kinase. Expression in multiple human tissues.
Multifunctional Ca2+/calmodulin-dependent protein kinase (CaM kinase) is a mediator of calcium signals in diverse signaling pathways. In human lymphocytes and epithelial tissues, CaM kinase activates a chloride channel via a Ca(2+)-dependent pathway which is preserved in cystic fibrosis. To characterize the CaM kinase present in these tissues we have cloned an isoform of this kinase from human T lymphocytes. We show the cDNA structure of two variants of this human CaM kinase, gamma B and gamma C, which are predicted to translate to 518 and 495 amino acids, respectively. Amino acid differences between these isoforms and the rat brain gamma isoform (which we refer to as gamma A) are localized to the variable domain. We used RNase protection of this variable region to reveal the level of expression of gamma B and gamma C CaM kinase mRNAs in nine human tissues and cell lines. When transfected into Jurkat T cells, the gamma B cDNA encoded a functional kinase which cosedimented on sucrose gradients with endogenous T cell CaM kinase activity and formed a large multimeric enzyme. The recombinant gamma B isoform displayed two phases of autophosphorylation characteristic of CaM kinases, including the phase which converts it to a partially Ca(2+)-independent species. Site-directed mutagenesis of the predicted autoinhibitory domain yielded a mutant which was approximately 37% active in the absence of Ca2+/calmodulin, confirming the region as critical for autoregulation, and suggesting this mutant as a tool for studying the role of CaM kinase in nonneuronal tissues.


Modelling vascular reactivity to investigate the basis of the relationship between cerebral blood volume and flow under CO2 manipulation.
Changes in cerebral blood flow (f) and vascular volume (v) are of major interest in mapping cerebral activity and metabolism, but the relation between them currently lacks a sufficient theoretical basis. To address this we considered three models: a uniform reactive tube model (M1); an extension of M1 that includes passive arterial inflow and venous volume (M2); and a more anatomically plausible model (M3) consisting of 19 compartments representing the whole range of vascular sizes and respective CO2 reactivities, derived from literature data. We find that M2 cannot be described as the simple scaling of a tube law, but any divergence from a linear approximation is negligible within the narrow physiological range encountered experimentally. In order to represent correctly the empirically observed slope of the overall v-f relationship, the reactive bed should constitute about half of the total vascular volume, thus including a significant fraction of capillaries and/or veins. Model M3 demonstrates systematic variation of the slope of the v-f relationship between 0.16 and 1.0, depending on the vascular compartment under consideration. This is further complicated when other experimental approaches such as flow velocity are used as substitute measurements. The effect is particularly large in microvascular compartments, but when averaged with larger vessels the variations in slope are contained within 0.25 to 0.55 under conditions typical for imaging methods. We conclude that the v-f relationship is not a fixed function but that both the shape and slope depend on the composition of the reactive volume and the experimental methods used.


Alteration of the erythrocyte ultrastructure and blood viscosity by morphine.
The effects of acute morphine administration on intact erythrocytes and on their flow properties were studied by measuring the mean cell volume, cell geometry, and whole blood and plasma viscosities. Morphine caused a small (2-7%) increase in mean cell volume. Changes in cell geometry were found to be time dependent and most pronounced in concave portions of the red cells. Whole body viscosity was found to decrease upon morphine treatment; this may be due in part to a concurrent decrease in plasma viscosity.


Rheumatologists' guideline adherence in rheumatoid arthritis: a randomised controlled study on electronic decision support, education and feedback.
To assess the effects of education, feedback and a computerised decision support system (CDSS) versus education and feedback alone on rheumatologists' rheumatoid arthritis (RA) guideline adherence. A single-centre, randomised controlled pilot study was performed among clinicians (rheumatologists, residents and physician assistants; n=20) working at the study centre, with a 1:1 randomisation of included clinicians. A standardized sum score (SSS) on guideline adherence was used as the primary outcome (patient level). The SSS was calculated from 13 dichotomous indicators on quality of RA monitoring, treatment and follow-up. The randomised controlled design was combined with a before-after design in the control group to assess the effect education and feedback alone. Twenty clinicians (mean age 44.3±10.9 years; 55% female) and 990 patients (mean age 62 ± 13 years; 69% female; 72% rheumatoid factor and/or anti-CCP positive) were included. Addition of CDSS to education and feedback did not result in significant better quality of RA care than education and feedback alone (SSS difference 0.02; 95%-CI -0.04 to 0.08; p=0.60). However, before/after comparison showed that education and feedback alone resulted in a significant increase in the SSS from 0.58 to 0.64 (difference 0.06; 95%-CI 0.02 to 0.11; p<0.01). Our results suggest that CDSS did not have added value with regard to guideline adherence, whereas education and feedback can lead to a small but significant improvement of guideline adherence.


First Report of Leaf Spot of Soybean Caused by Aristastoma guttulosum in China.
In fall, 2008, leaf spots were observed during the flowering stage of the Zhong Huang 13 cultivar of soybean in the fields of Anhui Province, China. The leaf spots were irregularly shaped, necrotic, brown-black, and surrounded by yellow halos. Often, on a given leaf, several spots joined one another to form a large blighted area. Finally, those leaves turned yellow followed by defoliation. Damaged leaves showed scattered black spots (i.e., numerous pycnidia) on the lower side. Fresh material was collected from infected plants and a single spore of the putative causal pathogen was isolated on potato dextrose agar (PDA) and incubated at 25°C during a 12-h dark/light cycle. The isolate produced a white fungal colony and black pycnidia after 30 days. The pycnidia are characterized as globose, dark brown-black, and distinctly papillate, with ostiolar setae, and are more or less straight, unbranched, and tapered at the apex. The conidia are clavate, hyaline, mostly with three transverse septa per cell; conidia are either straight or slightly bent, obviously guttulate, and 16 to 29 × 2.5 to 3.5 μm. This pathogen is similar to other Aristastoma guttulosum Sutton (1964), but with the following differences: (a) it has more than 10 versus 4 to 9 setae; (b) conidia are 16 to 29 × 2.5 to 3.5 μm versus 32 to 42 × 3.9 to 4.6 μm as reported for A. guttulosum (1). Conidia of the Chinese isolate were used to inoculate leaves of soybean. Five soybean leaves from potted plants, 1 month old, were sprayed with a suspension of conidia in water. Conidia were harvested from PDA cultures and the suspension was adjusted to 3 × 105 conidia/ml with a hemocytometer. Five leaves were sprayed with sterile distilled water as controls. Inoculated plants were kept in the greenhouse. All five of the inoculated leaves displayed the same symptoms observed in the fields. The symptoms developed initially as brown pinhead spots on the upper side of the leaves, gradually increasing to large brown spots. These spots were irregularly shaped, brown and necrotic in the center and surrounded by a yellow halo. Black pycnidia appeared after 1 week whereas the controls remained asymptomatic. The pathogen was reisolated from the inoculated soybean leaves according to standard Koch's postulates. Primers ITS1 and ITS4 were used in PCR reactions to amplify the internal transcribed spacer region (ITS) (3). Sequencing was performed using the same primers. The ITS sequence (GenBank Accession No. JF825548.1) for this pathogen (587 bp) was submitted to a BLAST search in GenBank. Since the ITS sequence of the genus Aristastoma has never been previously submitted, results did not show high similarity with any extant GenBank sequences. The genus Aristastoma Tehon (1933) was described by Tehon (2). Five of the species in this genus were described by Sutton (1). The number of septate conidium and lack of obvious guttulate within the conidium are the morphological basis to separate these five species. Morphological features of the pathogen from soybean leaves in China were slightly different from those of A. guttulosum. To our knowledge, this is the first report of leaf spot caused by A. guttulosum on soybean in China. References: (1) B. C. Sutton. Mycological Papers. 97:10, 1964. (2) L. R. Tehon. Mycologia XXV. 25:249, 1933. (3) T. J. White et al. Academic Press, San Diego, 1990.


Detecting binocular 3D motion in static 3D noise: no effect of viewing distance.
Relative binocular disparity cannot tell us the absolute 3D shape of an object, nor the 3D trajectory of its motion, unless the visual system has independent access to how far away the object is at any moment. Indeed, as the viewing distance is changed, the same disparate retinal motions will correspond to very different real 3D trajectories. In this paper we were interested in whether binocular 3D motion detection is affected by viewing distance. A visual search task was used, in which the observer is asked to detect a target dot, moving in 3D, amidst 3D stationary distractor dots. We found that distance does not affect detection performance. Motion-in-depth is consistently harder to detect than the equivalent lateral motion, for all viewing distances. For a constant retinal motion with both lateral and motion-in-depth components, detection performance is constant despite variations in viewing distance that produce large changes in the direction of the 3D trajectory. We conclude that binocular 3D motion detection relies on retinal, not absolute, visual signals.


Cutting edge: LPS-induced emergency myelopoiesis depends on TLR4-expressing nonhematopoietic cells.
Systemic bacterial infection is rapidly recognized as an emergency state leading to neutrophil release into the circulation and increased myeloid cell production within the bone marrow. However, the mechanisms of sensing infection and subsequent translation into emergency myelopoiesis have not been defined. In this study, we demonstrate in vivo in mice that, surprisingly, selective TLR4 expression within the hematopoietic compartment fails to induce LPS-driven emergency myelopoiesis. In contrast, TLR4-expressing nonhematopoietic cells are indispensable for LPS-induced, G-CSF-mediated myelopoietic responses. Furthermore, LPS-induced emergency myelopoiesis is independent of intact IL-1RI signaling and, thus, does not require inflammasome activation. Collectively, our findings reveal a key and nonredundant role for nonhematopoietic compartment pathogen sensing that is subsequently translated into cytokine release for enhanced, demand-adapted myeloid cell production.


Realizing the value in "non-standard" parts of the qPCR standard curve by integrating fundamentals of quantitative microbiology.
The real-time polymerase chain reaction (PCR), commonly known as quantitative PCR (qPCR), is increasingly common in environmental microbiology applications. During the COVID-19 pandemic, qPCR combined with reverse transcription (RT-qPCR) has been used to detect and quantify SARS-CoV-2 in clinical diagnoses and wastewater monitoring of local trends. Estimation of concentrations using qPCR often features a log-linear standard curve model calibrating quantification cycle (Cq) values obtained from underlying fluorescence measurements to standard concentrations. This process works well at high concentrations within a linear dynamic range but has diminishing reliability at low concentrations because it cannot explain "non-standard" data such as Cq values reflecting increasing variability at low concentrations or non-detects that do not yield Cq values at all. Here, fundamental probabilistic modeling concepts from classical quantitative microbiology were integrated into standard curve modeling approaches by reflecting well-understood mechanisms for random error in microbial data. This work showed that data diverging from the log-linear regression model at low concentrations as well as non-detects can be seamlessly integrated into enhanced standard curve analysis. The newly developed model provides improved representation of standard curve data at low concentrations while converging asymptotically upon conventional log-linear regression at high concentrations and adding no fitting parameters. Such modeling facilitates exploration of the effects of various random error mechanisms in experiments generating standard curve data, enables quantification of uncertainty in standard curve parameters, and is an important step toward quantifying uncertainty in qPCR-based concentration estimates. Improving understanding of the random error in qPCR data and standard curve modeling is especially important when low concentrations are of particular interest and inappropriate analysis can unduly affect interpretation, conclusions regarding lab performance, reported concentration estimates, and associated decision-making.


Twin kernel embedding.
In most existing dimensionality reduction algorithms, the main objective is to preserve relational structure among objects of the input space in a low dimensional embedding space. This is achieved by minimizing the inconsistency between two similarity/dissimilarity measures, one for the input data and the other for the embedded data, via a separate matching objective function. Based on this idea, a new dimensionality reduction method called Twin Kernel Embedding (TKE) is proposed. TKE addresses the problem of visualizing non-vectorial data that is difficult for conventional methods in practice due to the lack of efficient vectorial representation. TKE solves this problem by minimizing the inconsistency between the similarity measures captured respectively by their kernel Gram matrices in the two spaces. In the implementation, by optimizing a nonlinear objective function using the gradient descent algorithm, a local minimum can be reached. The results obtained include both the optimal similarity preserving embedding and the appropriate values for the hyperparameters of the kernel. Experimental evaluation on real non-vectorial datasets confirmed the effectiveness of TKE. TKE can be applied to other types of data beyond those mentioned in this paper whenever suitable measures of similarity/dissimilarity can be defined on the input data.


Persistence of SARS-CoV-2 antibodies over 18 months following infection: UK Biobank COVID-19 Serology Study.
Little is known about the persistence of antibodies after the first year following SARS-CoV-2 infection. We aimed to determine the proportion of individuals that maintain detectable levels of SARS-CoV-2 antibodies over an 18-month period following infection. Population-based prospective study of 20 000 UK Biobank participants and their adult relatives recruited in May 2020. The proportion of SARS-CoV-2 cases testing positive for immunoglobulin G (IgG) antibodies against the spike protein (IgG-S), and the nucleocapsid protein (IgG-N), was calculated at varying intervals following infection. Overall, 20 195 participants were recruited. Their median age was 56 years (IQR 39-68), 56% were female and 88% were of white ethnicity. The proportion of SARS-CoV-2 cases with IgG-S antibodies following infection remained high (92%, 95% CI 90%-93%) at 6 months after infection. Levels of IgG-N antibodies following infection gradually decreased from 92% (95% CI 88%-95%) at 3 months to 72% (95% CI 70%-75%) at 18 months. There was no strong evidence of heterogeneity in antibody persistence by age, sex, ethnicity or socioeconomic deprivation. This study adds to the limited evidence on the long-term persistence of antibodies following SARS-CoV-2 infection, with likely implications for waning immunity following infection and the use of IgG-N in population surveys.


Improving medication compliance: a randomised clinical trial.
A medical monitor which recorded the date and hour each time a medicine bottle was opened was used to evaluate a programme for improving patients' compliance with their treatment. Eighty-two patients with glaucoma who had been prescribed pilocarpine eye drops three times daily to prevent visual loss were randomised into two groups. Both groups used the medication monitor during two 20-day periods, but before the second period the experimental group were given an education and tailoring programme in an attempt to improve their compliance. Nine patients missed the second treatment period and were excluded from the analysis. The patients in the experimental group showed significantly improved compliance when compared with the control group. The numbers of missed doses were reduced by about half, as was the proportion of time that exceeded the eight-hour dose intervals. Follow-up studies are needed to determine how long the improved compliance persists, but anyone considering setting up an education and tailoring programme should recognise the extent to which therapeutic efforts are wasted because of non-compliance.


ProbeMaker: an extensible framework for design of sets of oligonucleotide probes.
Procedures for genetic analyses based on oligonucleotide probes are powerful tools that can allow highly parallel investigations of genetic material. Such procedures require the design of large sets of probes using application-specific design constraints. ProbeMaker is a software framework for computer-assisted design and analysis of sets of oligonucleotide probe sequences. The tool assists in the design of probes for sets of target sequences, incorporating sequence motifs for purposes such as amplification, visualization, or identification. An extension system allows the framework to be equipped with application-specific components for evaluation of probe sequences, and provides the possibility to include support for importing sequence data from a variety of file formats. ProbeMaker is a suitable tool for many different oligonucleotide design and analysis tasks, including the design of probe sets for various types of parallel genetic analyses, experimental validation of design parameters, and in silico testing of probe sequence evaluation algorithms.


Temporal construal and time-dependent changes in preference.
Five studies tested the predictions of temporal construal theory and time-discounting theories regarding evaluation of near future and distant future options (outcomes, activities, products). The options had abstract or goal-relevant features (called high-level construal features) as well as more concrete or goal-irrelevant features (called low-level construal features). The studies varied the valence (positive vs. negative) and the type of valence (affective vs. cognitive) of the low-level and high-level construal features. The results show that the weight of high-level construal features, compared with the weight of low-level construal features, is greater in determining distant future preferences than near future preferences. The implications of the results for extant theories of time-dependent changes in preference are discussed.


Structural investigations on nanoemulsions, solid lipid nanoparticles and nanostructured lipid carriers by cryo-field emission scanning electron microscopy and Raman spectroscopy.
Recently, colloidal dispersions based on solid lipids (solid lipid nanoparticles, SLN) and mixtures of solid and liquid lipids (nanostructured lipid carriers, NLC) were described as innovative carrier systems. A spherical particle shape is the basis of features such as a high loading capacity and controlled drug release characteristics due to smaller lipid-water interfaces and longer diffusion pathways when compared to thin platelets. The structures of SLN and the influence of oil load (NLC) on particle properties were investigated by photon correlation spectroscopy (PCS), laser diffractometry (LD), cryo-field emission scanning electron microscopy (cryo-FESEM), Raman spectroscopy and infrared spectroscopy (IR), and compared to a conventional nanoemulsion. PCS and LD data show similar size and size distribution for SLN and NLC (approximately 210 nm, polydispersity index approximately 0.15) and suggested a long term physical stability for the dispersions which had been stored for up to 12 months at different temperatures. Using cryo-FESEM droplets (for the nanoemulsion) and almost spherical particles for SLN and NLC were observed. Raman spectroscopy resulted in spectra for NLC that are weighted to the SLN spectra, suggesting an undisturbed crystal structure. Infrared spectra of the NLC are predominantly SLN in nature. Importantly the SLN bands are unshifted in the NLC spectrum indicating that the crystalline structure is unaffected by the presence of the oil.


Plasmon-modulated photoluminescence from gold nanostructures and its dependence on plasmon resonance, excitation energy, and band structure.
Two distinct single-photon plasmon-modulated photo-luminescence processes are generated from nanostructured gold surfaces by tuning the spectral overlap of the incident laser source, localized surface plasmon resonance band, and the interband transitions between the d and sp bands, near the X- and L-symmetry points of the electronic band structure of gold. In the main section of the article, the characteristics of these photoluminescence processes are described and discussed. In the last section, the background continuum accompanying surface-enhanced Raman scattering (SERS) spectra from benzenethiol and 4-mercaptopyridine self-assembled monolayers chemisorbed on nanostructured gold surfaces is shown to originate from plasmon-modulated photoluminescence.


Collective Impact Approaches to Promoting Community Health and Wellbeing in a Regional Township: Learnings for Integrated Care.
This Perspective Paper explores the challenges of implementing local initiatives guided by the tenets of the Collective Impact (CI) approach. As such, it draws implications of CI for integrated health and social care efforts to improve and sustain health and social outcomes within a community-wide context, based on our efforts to deploy a CI intervention in the regional town of Muswellbrook, New South Wales (NSW) Australia. A program of health and wellbeing activities providing mental health and wellness messages and activities was implemented in the township over 2 years by the Family Action Centre (FAC), University of Newcastle, Australia. A key takeaway was the importance of authentic community engagement and active involvement as opposed to mere consultation.


The IFN-γ receptor promotes immune dysregulation and disease in STING gain-of-function mice.
STING gain-of-function mutations cause STING-associated vasculopathy with onset in infancy (SAVI) in humans, a disease characterized by spontaneous lung inflammation and fibrosis. Mice with STING gain-of-function mutations (SAVI mice) develop αβ T cell-dependent lung disease and also lack lymph nodes. Although SAVI has been regarded as a type I interferonopathy, the relative contributions of the three interferon receptors are incompletely understood. Here, we show that STING gain of function led to upregulation of IFN-γ-induced chemokines in the lungs of SAVI mice and that deletion of the type II IFN receptor (IFNGR1), but not the type I IFN receptor (IFNAR1) or type III IFN receptor (IFNλR1), ameliorated lung disease and restored lymph node development in SAVI mice. Furthermore, deletion of IFNGR1, but not IFNAR1 or IFNλR1, corrected the ratio of effector to Tregs in SAVI mice and in mixed bone marrow chimeric mice. Finally, cultured SAVI mouse macrophages were hyperresponsive to IFN-γ, but not IFN-β, in terms of Cxcl9 upregulation and cell activation. These results demonstrate that IFNGR1 plays a major role in autoinflammation and immune dysregulation mediated by STING gain of function.


Trait-state artifacts and the diagnosis of personality disorders.
The multiaxial nature of DSM-III has stimulated interest in the personality disorders. There are also indications that it has produced an increase in their diagnosis. However, there is clinical and psychometric evidence that a personality evaluation undertaken while a patient is in a dysphoric mental state may distort or misrepresent traits, the so-called trait-state problem in personality assessment. The present study appears to be the first to investigate this phenomenon with a clinical interview rather than with personality tests. It examined the effect of anxiety, depression, and level of global impairment on the diagnosis of personality disorder and the assessment of the criteria for the individual Axis II disorders. Eighty-four patients, most of whom had current Axis I diagnoses, were evaluated by seven experienced clinicians with a new semistructured interview, the Personality Disorder Examination. The sample evidenced a trend toward acknowledging fewer maladaptive personality traits at follow-up than at entry. There was no evidence, however, that anxiety or depression had affected either the diagnosis of a personality disorder or the criteria associated with most of the individual personality disorders.


An in vivo study on brain microstructure in biological and chronological ageing.
This study aimed to investigate whether magnetization transfer imaging (MTI) parameters of cortical gray and white matter and subcortical gray matter structures differ between subjects enriched for human familial longevity and control subjects to provide a thorough description of the brain phenotype of familial longevity. Moreover, we aimed to describe cerebral ageing effects on MTI parameters in an elderly cohort. All subjects were included from the Leiden Longevity Study and underwent 3 Tesla MTI of the brain. In total, 183 offspring of nonagenarian siblings, who are enriched for familial factors of longevity, were contrasted with 163 environmentally and age-matched controls. No differences in cortical and subcortical gray matter and white matter MTI parameters were found between offspring and control subjects using histogram-based and voxel-wise analyses. Cortical gray matter and white matter MTI parameters decreased with increasing chronological age (all p < 0.001). Decrease of white matter magnetization transfer ratio (MTR) was homogeneous throughout the whole mean white matter skeleton except for parts of the callosal splenium and partly the posterior limb of the internal capsule and superior region of the corona radiata (p < 0.05). Mean MTR of subcortical gray matter structures decreased with increasing age (p amygdala, caudate nucleus and putamen < 0.001; p pallidum = 0.001, p thalamus = 0.002). In conclusion, the brain phenotype of human familial longevity is - at a mean age of 66 years - not characterized by preserved macromolecular brain tissue integrity.


Factors affecting wound leakage in 23-gauge sutureless pars plana vitrectomy.
To evaluate risk factors for sclerotomy leakage in 23-gauge sutureless pars plana vitrectomy in 219 patients. Nested case-control study involving 48 patients with wound leaks (visible on-table sclerotomy leakage requiring sutures) and 171 control subjects without wound leaks. Patients received either a conventional sclerotomy incision at 45°, which was then changed to 90° midincision, or an extremely oblique sclerotomy incision (OSI) at 10°, which was then changed to 30° midincision. Risk factors studied included age, gender, laterality, surgical duration, sclerotomy incision (OSI vs. conventional sclerotomy incision), preoperative diagnosis (macular vs. nonmacular), history of vitrectomy, and primary surgeon (attending vs. supervised resident). Multivariate logistic regression analysis found significant (P ≤ 0.05) protective factors for wound leakage including OSI, macular preoperative diagnosis, no previous vitrectomy, and female gender. Surgical duration at least 45 minutes was considered a borderline risk factor. Using an extremely OSI versus a conventional sclerotomy incision reduces the incidence of wound leakage postoperatively because of its self-sealing effect. Other factors that contribute to wound leakage, such as increased surgical duration and nonmacular diagnosis, may be indirect measurements of extensive trocar rotation, causing wound leakage despite the use of an OSI.


In vivo performance of selective electron beam-melted Ti-6Al-4V structures.
Highly porous titanium structures are widely used for maxillofacial and orthopedic surgery because of their excellent mechanical properties similar to those of human bone and their facilitation of bone ingrowth. In contrast to common methods, the generation of porous titaniumproducts by selective electron beam melting (SEBM), an additive manufacturing technology, overcomes difficulties concerning the extreme chemical affinity of liquid titanium to atmospheric gases which consequently leads to strongly reduced ductility of the metal. The purpose of this study was to assess the suitability of a smooth compact and a porous Ti-6Al-4V structure directly produced by the SEBM process as scaffolds for bone formation. SEBM-processed titanium implants were placed into defects in the frontal skull of 15 domestic pigs. To evaluate the direct contact between bone and implant surfaces and to assess the ingrowth of osseous tissue into the porous structure, microradiographs and histomorphometric analyses were performed 14, 30, and 60 days after surgery. Bone ingrowth increased significantly during the period of this study. After 14 days the most outer regions of the implants were already filled with newly formed bone tissue (around 14%). After 30 days the bone volume inside the implants reached almost 30% and after 60 days abundant bone formation inside the implants attained 46%. During the study only scarce bone-implant contact was found around all implants, which did not exceed 9% around compact specimens and 6% around porous specimens after 60 days. This work demonstrates that highly porous titanium implants with excellent interconnectivity manufactured using the SEBM method are suitable scaffolds for bone ingrowth. This technique is a good candidate for orthopedic and maxillofacial applications.


Using the PhenX Toolkit to Add Standard Measures to a Study.
The PhenX (consensus measures for Phenotypes and eXposures) Toolkit (https://www.phenxtoolkit.org/) offers high-quality, well-established measures of phenotypes and exposures for use by the scientific community. The goal is to promote the use of standard measures, enhance data interoperability, and help investigators identify opportunities for collaborative and translational research. The Toolkit contains 395 measures drawn from 22 research domains (fields of research), along with additional collections of measures for Substance Abuse and Addiction (SAA) research, Mental Health Research (MHR), and Tobacco Regulatory Research (TRR). Additional measures for TRR that are expected to be released in 2015 include Obesity, Eating Disorders, and Sickle Cell Disease. Measures are selected by working groups of domain experts using a consensus process that includes input from the scientific community. The Toolkit provides a description of each PhenX measure, the rationale for including it in the Toolkit, protocol(s) for collecting the measure, and supporting documentation. Users can browse measures in the Toolkit or can search the Toolkit using the Smart Query Tool or a full text search. PhenX Toolkit users select measures of interest to add to their Toolkit. Registered Toolkit users can save their Toolkit and return to it later to revise or complete. They then have options to download a customized Data Collection Worksheet that specifies the data to be collected, and a Data Dictionary that describes each variable included in the Data Collection Worksheet. The Toolkit also has a Register Your Study feature that facilitates cross-study collaboration by allowing users to find other investigators using the same PhenX measures.


Biodemography of human ageing.
Human senescence has been delayed by a decade. This finding, documented in 1994 and bolstered since, is a fundamental discovery about the biology of human ageing, and one with profound implications for individuals, society and the economy. Remarkably, the rate of deterioration with age seems to be constant across individuals and over time: it seems that death is being delayed because people are reaching old age in better health. Research by demographers, epidemiologists and other biomedical researchers suggests that further progress is likely to be made in advancing the frontier of survival - and healthy survival - to even greater ages.


Menopausal bone changes and incident fractures in diabetic women: a cohort study.
The purpose of this study was to evaluate the rate of bone loss and incident fractures in women with diabetes mellitus (DM) across menopause. During menopause, DM women experienced bone mineral density (BMD) loss that was faster at hip and slower at spine and had a higher risk of fractures, perhaps because of their earlier menopause. The increasing DM epidemic will contribute to higher fracture burden. Women with DM have a higher risk of fractures independent of age, body mass index (BMI), and BMD. Our objective is to evaluate if women with DM experience greater bone loss and more fractures across menopause. Two thousand one hundred seventy one women, aged 42 to 52 years at baseline (1996), enrolled in the Study of Women's Health Across the Nation (SWAN), a prospective study, with 8 years of annual follow up. One thousand three hundred forty six (62%) completed annual visit 7 (2004). Women with baseline fasting blood glucose level of ≥126 mg/dl and those being treated for diabetes were designated as DM. Annual assessment of menopausal stage, BMD, and urinary N-telopeptide (NTx) were carried out. Rate of change in BMD across menopause and annual self-report data for risk of incident fractures by DM status were determined. Despite higher baseline BMD at hip (p = <0.001), and lumbar spine (p = <0.001), rate of decline in BMD was faster at hip (β = -0.45 vs. -0.11 gm/cm(2)/year, p = <0.001) for DM women, compared to non-DM. However, lumbar spine bone loss was slower in women with DM as compared to non-DM women (β = 0.04 vs. -0.25 gm/cm(2)/year, p = 0.004). DM women experienced menopause 3 years earlier than non-DM women (p = 0.002), and age adjusted incident fractures were two fold higher in women with DM compared to non-DM (RR = 2.20, 95% CI: 1.26-3.85, p = <0.006). BMD loss is greater in hip and slower at spine in DM women during menopausal transition. Women with DM have a higher risk of fractures, perhaps because of their earlier menopause.


Drug interactions with the tyrosine kinase inhibitors imatinib, dasatinib, and nilotinib.
Several cancer treatments are shifting from traditional, time-limited, nonspecific cytotoxic chemotherapy cycles to continuous oral treatment with specific protein-targeted therapies. In this line, imatinib mesylate, a selective tyrosine kinases inhibitor (TKI), has excellent efficacy in the treatment of chronic myeloid leukemia. It has opened the way to the development of additional TKIs against chronic myeloid leukemia, including nilotinib and dasatinib. TKIs are prescribed for prolonged periods, often in patients with comorbidities. Therefore, they are regularly co-administered along with treatments at risk of drug-drug interactions. This aspect has been partially addressed so far, calling for a comprehensive review of the published data. We review here the available evidence and pharmacologic mechanisms of interactions between imatinib, dasatinib, and nilotinib and widely prescribed co-medications, including known inhibitors or inducers of cytochromes P450 or drug transporters. Information is mostly available for imatinib mesylate, well introduced in clinical practice. Several pharmacokinetic aspects yet remain insufficiently investigated for these drugs. Regular updates will be mandatory and so is the prospective reporting of unexpected clinical observations.


Mesenrhombencephalitis: MR findings in nine patients.
Mesenrhombencephalitis is a serious form of brainstem inflammation predominantly involving the deep and vital portions of the brain, that is, the mesencephalon (midbrain) and rhombencephalon (pons, medulla). Mesenrhombencephalitis is difficult to diagnose on the basis of clinical and laboratory findings alone, and access to this portion of the brain for surgical biopsy carries high morbidity. We describe the MR appearance of mesenrhombencephalitis and correlate the imaging findings with clinical information. Unenhanced and contrast-enhanced MR images of nine patients with mesenrhombencephalitis were reviewed retrospectively and correlated with clinical, laboratory, and pathologic data. The patients were categorized according to the cause of the disease: three had herpes simplex, one had Listeria monocytogenes, and five had mesenrhombencephalitis of undetermined cause. The three patients with clinical and MR evidence of herpes simplex mesenrhombencephalitis (one confirmed by brain biopsy) were comatose at presentation, with cranial nerve abnormalities in two and seizures in one. One patient with L. monocytogenes (established by blood culture) had cranial nerve palsies, fever, and pain in the ear. Five additional patients had headache (three), fever (three), nausea and vomiting (four), cranial nerve palsies (three), coma (two), and hyporeflexia (one) or hyperreflexia (four). Brain biopsy performed in two patients revealed chronic inflammation of unspecified cause; in one, it was compatible with viral encephalitis. MR images in three patients with herpes simplex mesenrhombencephalitis showed T2 signal hyperintensity in the midbrain (two), pons (one), medulla (one), and temporal lobes (three). Parenchymal foci of hemorrhage (methemoglobin, one patient) and leptomeningeal enhancement (one patient) were identified in the temporal lobes. T2-weighted MR images in one patient with L. monocytogenes showed signal hyperintensity in the brainstem, vermis, midbrain, and internal capsules. On T1-weighted images, low signal was present in these areas, which enhanced with paramagnetic contrast agents. In the remaining five patients, T2-weighted MR images showed patchy signal hyperintensity in the pons, medulla, and thalamus in three each and in the midbrain and temporal lobes in one each. T1-weighted MR images showed normal findings (two) or signal hypointensity in the thalamus and pons in one patient each. Areas of leptomeningeal and parenchymal enhancement were identified in one patient each. Brainstem swelling was seen in three patients, one of whom had petechial hemorrhage in the pons and hydrocephalus. Mesenrhombencephalitis is a serious illness that is diagnosed by a combination of imaging, clinical, laboratory, and pathologic studies. MR imaging may be crucial to the early diagnosis of this illness, and radiologists must be familiar with this uncommon entity and its MR findings in order to make timely diagnoses and facilitate treatment.


Independent and interdependent functions of LAF1 and HFR1 in phytochrome A signaling.
Several positive regulators of phytochrome A signaling--e.g., LAF1, HFR1, and HY5--operate downstream from the photoreceptor, but their relative sites of action in the transduction pathway are unknown. Here, we show that HFR1RNAi/laf1 or hfr1-201/LAF1RNAi generated by RNA interference (RNAi) has an additive phenotype under FR light compared with the single mutants, hfr1-201 or laf1. This result indicates that LAF1 and HFR1 function in largely independent pathways. LAF1, an R2R3-MYB factor, interacts with HFR1, a basic helix-loop-helix (bHLH) factor, and this interaction is abolished by the R97A mutation in the LAF1 R2R3 domain. Polyubiquitinations of LAF1 and HFR1 by the COP1 E3 ligase in vitro are inhibited by LAF1/HFR1 association. Consistent with this result, endogenous HFR1 is less stable in laf1 compared with wild type, and similarly, LAF1-3HA expressed from a transgene is also less stable in hfr1-201 than wild type. In transgenic plants, HFR1 levels are significantly elevated upon induced expression of LAF1 but not LAF1(R97A). Moreover, induced expression of LAF1 but not LAF1(R97A) delays post-translational HFR1 degradation in FR light. Constitutive coexpression of HFR1 and LAF1 but not HFR1 and LAF1 (R97A) confers FR hypersensitivity in double transgenic plants. Our results show that in addition to their independent functions in phyA signaling, LAF1 and HFR1 also cooperate post-translationally to stabilize each other through inhibition of ubiquitination by COP1, thereby enhancing phyA photoresponses.


Patterns of illness explaining the associations between posttraumatic stress disorder and the use of CT.
To examine the relationship between posttraumatic stress disorder (PTSD) and computed tomography (CT) utilization and to determine whether there were patterns of comorbid illness that could explain the relationship. The study was approved by the University of Iowa Institutional Review Board and the Iowa City Veterans Affairs Medical Center Research and Development Committee. By using a retrospective cohort design, a national sample of new veteran enrollees aged 18-35 years was studied. Associations were examined between the presence of PTSD, receipt of at least one and multiple CT scans, comorbid medical conditions (eg, abdominal pain, headaches), and measures of health care utilization (eg, primary care, emergency room, and mental health visits) and the daily probability of the receipt of at least one CT scan before and after a diagnosis of PTSD. Analyses included sequential multivariable generalized linear mixed models to examine the independent relationship between PTSD and CT scan utilization. Among the full cohort, 13.0% (10 018 of 76 812) received at least one CT scan. PTSD was identified in 21.1% (16 182 of 76 812) of the cohort, and 22.9% (3711 of 16 182) of veterans with PTSD received at least one CT scan as compared with 10.4% (6307 of 60 630) of veterans without PTSD (P < .0001). In sequential modeling, comorbid factors explaining the relationship between CT scans and PTSD were traumatic brain injury (odds ratio, 3.54; P < .0001), abdominal pain (odds ratio, 4.01; P < .0001), and headaches (odds ratio, 3.07; P < .0001). Associations were also strong for high levels of emergency room (odds ratio, 2.73; P < .0001) and primary care (odds ratio, 2.38; P < .0001) utilization. The daily chance of receiving a CT scan was seven times higher prior to the recognition of PTSD (daily chance, 0.007 before vs 0.001 after; P < .0001). Young veterans with PTSD are receiving more CT scans compared with those without PTSD; the daily probability of receiving CT scans is higher prior to recognition of PTSD. http://radiology.rsna.org/lookup/suppl/doi:10.1148/radiol.13121593/-/DC1.


Use of over-the-counter malaria medicines in children and adults in three districts in Kenya: implications for private medicine retailer interventions.
Global malaria control strategies highlight the need to increase early uptake of effective antimalarials for childhood fevers in endemic settings, based on a presumptive diagnosis of malaria in this age group. Many control programmes identify private medicine sellers as important targets to promote effective early treatment, based on reported widespread inadequate childhood fever treatment practices involving the retail sector. Data on adult use of over-the-counter (OTC) medicines is limited. This study aimed to assess childhood and adult patterns of OTC medicine use to inform national medicine retailer programmes in Kenya and other similar settings. Large-scale cluster randomized surveys of treatment seeking practices and malaria parasite prevalence were conducted for recent fevers in children under five years and recent acute illnesses in adults in three districts in Kenya with differing malaria endemicity. A total of 12, 445 households were visited and data collected on recent illnesses in 11, 505 children and 19, 914 adults. OTC medicines were the most popular first response to fever in children with fever (47.0%; 95% CI 45.5, 48.5) and adults with acute illnesses (56.8%; 95% CI 55.2, 58.3). 36.9% (95% CI 34.7, 39.2) adults and 22.7% (95% CI 20.9, 24.6) children using OTC medicines purchased antimalarials, with similar proportions in low and high endemicity districts. 1.9% (95% CI 0.8, 4.2) adults and 12.1% (95% CI 16.3,34.2) children used multidose antimalarials appropriately. Although the majority of children and adults sought no further treatment, self-referral to a health facility within 72 hours of illness onset was the commonest pattern amongst those seeking further help. In these surveys, OTC medicines were popular first treatments for fever in children or acute illnesses in adults. The proportions using OTC antimalarials were similar in areas of high and low malaria endemicity. In all districts, adults were more likely to self-treat with OTC antimalarial medicines than febrile children were to receive them, and less likely to use them in recommended ways. Government health centres were the most common second resort for treatment and were often used within 72 hours. In view of these practices, more research is needed to assess the impact on the popularity of private medicine sellers of strengthened public sector policies on access to malaria treatment and insecticide-treated bed nets. Improved targeting of OTC antimalarials to high risk groups, better communication strategies regarding adult as well as children's dosages, and facilitating more rapid referral to trained health workers where needed are important challenges to private medicine seller programmes.